A phage-displayed chicken single-chain antibody fused to alkaline phosphatase detects Fusarium pathogens and their presence in cereal grains
文献类型: 外文期刊
第一作者: Hu, Zu-Quan
作者: Hu, Zu-Quan;Li, He-Ping;Zhang, Jing-Bo;Huang, Tao;Liu, Jin-Long;Xue, Sheng;Liao, Yu-Cai;Li, He-Ping;Huang, Tao;Zhang, Jing-Bo;Liao, Yu-Cai;Wu, Ai-Bo;Liao, Yu-Cai
作者机构:
关键词: Fusarium pathogens;Single-chain variable fragment;Phage display;Alkaline phosphatase;Fusion protein;Enzyme-linked immunosorbent assay
期刊名称:ANALYTICA CHIMICA ACTA ( 影响因子:6.558; 五年影响因子:6.228 )
ISSN:
年卷期:
页码:
收录情况: SCI
摘要: Fusarium and its poisonous mycotoxins are distributed worldwide and are of particular interest in agriculture and food safety. A simple analytical method to detect pathogens is essential for forecasting diseases and controlling mycotoxins. This article describes a proposed method for convenient and sensitive detection of Fusarium pathogens that uses the fusion of single-chain variable fragment (scFv) and alkaline phosphatase (AP). A highly reactive scFv antibody specific to soluble cell wall-bound proteins (SCWPs) of F. verticillioides was selected from an immunized chicken phagemid library by phage display. The antibody was verified to bind on the surface of ungerminated conidiospores and mycelia of F. verticillioides. The scFv-AP fusion was constructed, and soluble expression in bacteria was confirmed. Both the antibody properties and enzymatic activity were retained, and the antigen-binding capacity of the fusion was enhanced by the addition of a linker. Surface plasmon resonance measurements confirmed that the fusion displayed 4-fold higher affinity compared with the fusion's parental scFv antibody. Immunoblot analyses showed that the fusion had good binding capacity to the components from SCWPs of F. verticillioides, and enzyme-linked immunosorbent assays revealed that the detection limit of the fungus was below
分类号: O65
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